Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Immunoglobulin molecules with improved characteristics

a technology of immunoglobulin and characteristics, applied in the field of immunology and molecular biology, to achieve the effect of increasing the serum half life of an igg1

Active Publication Date: 2012-04-24
BOARD OF RGT THE UNIV OF TEXAS SYST
View PDF6 Cites 104 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

"The present invention provides an IgG1 molecule that can bind to a pathogen antigen, a toxin, an autoimmune antibody, or an anti-transplant antibody. The molecule can also be used to block the function of FcRn, a protein that plays a role in the immune system. The molecule can be administered to a subject to treat or prevent infections or autoimmune disorders. The molecule has a specific structure and can be produced in a pharmaceutical composition. The invention also provides a method of delivering the molecule to a fetus and a method of blocking FcRn function in a subject."

Problems solved by technology

If IgG homesostasis is disturbed, then pathology due to too high (hypergammaglobunemia) or too low (hypogammaglobunemia) can result.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Immunoglobulin molecules with improved characteristics
  • Immunoglobulin molecules with improved characteristics
  • Immunoglobulin molecules with improved characteristics

Examples

Experimental program
Comparison scheme
Effect test

example 1

Materials & Methods

[0086]Site directed mutagenesis. The human IgG1 constant regions were mutated using the PCR with mutagenic oligonucleotides and splicing by overlap extension (Horton et al., 1989). Restriction sites encompassing the mutated sequence in the constant region gene were used for recloning the mutated regions as smaller subfragments of the entire constant region gene. Following recloning, constant region genes were sequenced to ensure insertion of the desired mutation without second site mutations. Mutated genes were recloned into the final expression construct (FIG. 2) using standard methods of molecular biology.

[0087]Transfection for expression of antibodies. NSO cells expressing a human anti-lysozyme specific light chain (HuLys5: Foote and Winter, 1992) were transfected by electroporation with heavy chain expression constructs (FIG. 2) encoding an anti-lysozyme heavy chain with mutations in the Fc region as described. Transfectants were selected in medium containing ...

example 2

Results

[0093]Analyses of the interactions of human FcRn with wild-type human IgG1 and human IgG1 mutants containing Lys433, Phe434 and His436 (mutated to corresponding mouse IgG1 residue at position 436) or Lys433, Phe434 and Tyr436 (wild-type residue at position 436) indicates that the mutant with the wild-type Tyr436 has an approximately two-fold higher affinity than the mutant containing His436 (Table I). In addition, both mutants have substantially higher affinities for human FcRn than the corresponding wild-type human IgG1. These affinities are further improved when mutations of Met252 to Tyr, Ser254 to Thr and Thr256 to Glu are added to the position 433, 434 (and 436) mutations. Higher affinity for binding to human FcRn is retained for the human IgG1 variant containing the wild-type residue Tyr, rather than histidine, at position 436 (Table II). Taken together, the data therefore indicate that Tyr436 is preferred over His436 for the interaction of human IgG1 with human FcRn.

[0...

example 3

Methods & Results

[0100]Labeling of antibodies. Antibodies were labeled using lodogen or biotinylated in 0.1 M carbonate buffer (pH 8.5) using 9 μg EZ-link Sulfo-NHS biotin (Pierce) per milligram of antibody as described (Firan et al., 2001). Prior to use in studies, all labeled antibodies were compared with their unlabeled counterparts using surface plasmon resonance to ensure that labeling had not altered their binding characteristics.

[0101]Placental transport assays. Essentially the same methods as those described previously were used (Firan et al., 2001). Three mgs of each of wild-type IgG1 or human IgG1 mutant containing Lys433 and Phe434 with the wild-type residue Tyr at position 436 (labeled with either biotin or 125I) were added to the maternal compartment of the ex vivo placental model and transport to the fetal compartment analyzed by sample collection at the indicated times. Following collection, samples were centrifuged at 1500 g for 10 minutes and pellets discarded. The ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
pHaaaaaaaaaa
flow rateaaaaaaaaaa
concentrationsaaaaaaaaaa
Login to View More

Abstract

The present invention provides for IgG1 molecules with improved characteristics. In particular, substitution mutations are provided that, in combination, facilitate improved placental transfer, improved serum half-life and improved FcRn binding. Substitution mutations are also provided, that in combination, can be used to block FcRn function and thereby increase the clearance rates of other (endogenous or exogenous) IgGs, block placental transport of IgGs and have increased affinity / reduced pH dependence for FcRn binding.

Description

[0001]This application claims benefit of priority to U.S. Provisional Application Ser. No. 60 / 685,934, filed May 31, 2005, the entire content of which is hereby incorporated by reference.[0002]This invention was made with government support under grant numbers R01 AI39167 and R01 AI55556 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND OF THE INVENTION[0003]1. Field of the Invention[0004]The present invention relates generally to the fields of immunology and molecular biology. More specifically, the invention relates to modified Ig molecules with improved characteristics.[0005]2. Description of the Related Art[0006]Immunoglobulin G (IgG) constitutes the most prevalent immunoglobin class in the serum of man and other mammals. Despite fluctuations in rates of synthesis by B cells, IgGs are maintained at remarkably constant levels in the serum. If IgG homesostasis is disturbed, then pathology due to too high (hypergammaglobunem...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(United States)
IPC IPC(8): C07K16/00C12P21/08C07K1/00
CPCA61K47/48384A61K49/0058C07K16/00C07K16/40A61K2039/505C07K2317/21C07K2317/92C07K2317/52A61K47/6803A61P19/04A61P21/04A61P29/00A61P35/00A61P35/02A61P35/04A61P37/00A61P37/06A61P5/16A61K51/1075
Inventor OBER, E. SALLY WARD
Owner BOARD OF RGT THE UNIV OF TEXAS SYST
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com